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Practical 4: H&E
Dr Louise Dunford
Learning outcomes
Identify skin & ileum sections
Describe the different regions/structures
within your stained section
Review slides for quality of staining
Understand how staining can be improved
Skin
Histology video of thin skin
http://www.youtube.com/watch?
v=KmfEbKmKFw
Skin
Boston
University
Skin
Boston
University
Skin
Boston
University
Skin
Boston
University
GI Tract
Ileum
Histology video of the ileum:
https://
www.youtube.com/watch?v=M6
zcmsy8c-o
Ileum
Boston University
Peyers patches
Ileum
Boston
University
Ileum
Boston
University
Haematoxylin
Most staining is a simple nuclear stain and a
counterstain
In most staining protocols the lowest molecular weight
dyes are applied first (e.g. haematoxylin), differentiated,
and then larger counterstains are applied
Haematoxylin is a mordant (metal complexing) dye; the
cationic/basic dye-metal complex binds to nucleic acids
in DNA
Eosin
Anionic/acid counterstain
Eosin Y binds to amino acids and most cellular
components, but not for example, larger
molecular structures such as collagen which
are also anionic
Stains pink/red
Troubleshooting
All stages need to be carried out in optimal
conditions:
Collection of the tissue
Fixation
Processing
Cutting
Troubleshooting
Troubleshooting
Troubleshooting
Troubleshooting
Troubleshooting
Troubleshooting
Troubleshooting
Troubleshooting
Resources
University of Leeds Histology website:
http://www.histology.leeds.ac.uk/skin/index.php
Young, B et al., (2006)., Wheaters Functional Histology:
A Text and Colour Atlas. Churchill Livingstone Elsevier
A troubleshooting guide:
http://
www.leica-microsystems.com/uploads/media/101_steps_to
_better_histology.pdf
Learning outcomes
Identify skin & ileum sections
Describe the different regions/structures
within your stained section
Review slides for quality of staining
Understand how staining can be improved